II‘ I “ I ‘4 II III‘ III I I IIII I I I I I I IIIIIIII I I l I I I I I I II I I I I IIIIIIIIIII (”—3 I 00 I (DA II INFLUENCE OF MEDIUM AND HIGH PROTEIN DIETS ON THE CHLORINE METABOLISM OF TWO PRESCHOOL. r ”CHILDREN f _ ’ THESIS FOR THE DEGREE III III s _ Mar-garct Bérry 1933‘; IIIII III III 72 IIIIIII 01 III III 9 III I III ‘(cmaa 7;, gawnvhtf'lllllll I. . I shirt I . I llallilivlrivhklfkbvlr ,DLil I I I. [9.53.2 ITIEME.’.I I I .IESIE'FIIIIPKI. I. und; w IIquIVWKLH’lu I}?! lefiux‘Imuw H BI III-Isl 11.4 4.. Ha. Tr ‘ INFLUENCE OF MEDIUM AND HIGH PROTEIN DIETS ‘ ON THE CHLORIDE METABOLISM OF TWO PRESCHOOL CHILDREN by. / gwa,a., Harraret Barry / / 5 fl . / L! A Thesis Submitted to the Faculty of Michigan State College in Partial Fulfillment of the Requirements for the Degree of Master of Science Department of Foods and Nutrition Division of Home Economics 1933 ACKNOWLEDGMENT I wish to eXpress my appreciation to Dr. Marie Dye and Dr. Jean Hawks for their helpful criticisms throughout this study. 93855 TKBLE OF CONTENTS List of Tables Introduction Review of Literature Procedure Discussion of Results Summary BibliOgraphy Page 12 18 35 Table I. II. III. IV. v. VI. VII. VIII. IX. XI. XII. XIII. XIV. LIST OF TABLES Page Data on the Chloride Metabolism of Healthy 3 Infants Summary of the Data on Chlorine Metabolism as Reported by Various Authors 5 Summary of Clark's Chlorine Metabolism Studies on Men 10 The Height and Height of the Children as Compared to the Woodbury Height-Weight—Age Tables 12 Composition of the Medium Protein Diet 1n Composition of the High Protein Diet 15 Results of Tests to Show the Accuracy of the Chlorine Method 17 Daily Chlorine Metabolism on a Medium Protein Diet 19 Three Day Average of the Daily Chlorine Metabolism on a Medium Protein Diet 20 The Daily Chlorine Metabolism on a Medium Protein Diet Expressed in Grams per KilOgram of Body Weight 23 Three Day Average of the Daily Chlorine Metabolism on a Medium Protein Diet Expressed in Grams per Kilogram of Body Weight 25 Daily Chlorine Metabolism on a High Protein Diet 26 Three Day Average of the Daily Chlorine Metabolism, on a lfiigh. Protein Diet 28 Daily Chlorine Metabolism on a High Protein Diet EXpressed in Grams per KiIOgram of Body Weight 31 List of Tables (Cont'd) Table Page XV. Three Day Average of the Daily Chlorine metabolism on a High Protein Diet Ex— pressed in Grams per Kilogram of Body Weight 32 Appendix: XVI. Derily Chlorine Metabolism on a Medium Protein Diet EXpressed in Grams of Sodium Chloride 37 XVII. The Daily Chlorine Metabolism on a High Protein Diet EXpressed in Grams of Sodium Chloride 38 INFLUEHCE O. MEDIUM AND HIGH PROTEIN DIETS ON THE CHLOHIHE HETABOLISH OF TWO PRESCHOOL CHILDREN Lgtggduction It has long been known that chlorides are important in many pathological conditions and especially in the treatment of the edema of nephritis. This is true, because of the ability of chlorides to regulate the fluid content of the body. Chlorides also help to maintain the physical neutrality in the body fluids, and are significant in conditions in which there is a disturbance of the acid-base equilibrium. Another function of the chlorides is to aid in the control of the relative distribution of water between the circulating blood and body cells. However, the knowledge is meager regarding the chlorine needs of the normal in- dividual, and additional study of this element in respect to chlorine metabolism should help to further determine its function. The objects of this investigation were first, to deter- mine the utilization of chlorine by two normal, preschool children receiving weighed diets of either medium or high protein content, second, to compare the chlorine metabolism of the two children, third, to determine to some extent the the daily variations in the chlorine metabolism of the two subjects, and fourth, to give some information about the amount of chlorine used by children of this age. Review 9; Literature A review of American literature revealed very few studies concerning the chlorine metabolism of subjects of any age. A few investigators have studied the chlorine metabolism of infants fed various kinds of milk. Wang and Davis (12) reviewed the work done by foreign scientists and their results are summarized in Table I. Observation of this table reveals that, with three exceptions, the amount of chlorine ingested on the basis of gram per kilogram of body weight varied from 0.03% to 0.081 gram for the different subjects. Although the percentage of the intake absorbed varied from 59 to 97 percent, the amount absorbed per kilo- gram of body weight was quite similar in the majority of cases, and varied directly with the amount ingested. The chlorine retention fluctuated from 8 to Sh percent of the intake. The amount retained, however, was surprisingly con— stant, ten out of the twelve balances reported varying only between 0.011 and 0.027 gram per kilogram of body weight. Tangle fed a fairly high chlorine intake to his two cases and observed that only 8 percent of the intake was retained. These retentions were low when compared to those ranging from 20 to 50 percent of the intake, as recorded by other invest— igators. In their own study, Wang and Davis (12) compared the metabolism of the mineral constituents of cows milk and breast milk on five apparently healthy infants between the ages of -2... F TABLE I DATA ON THE CHLORIDE 2201801182 OF HEALTH" INFANTS* _ Subject Length Chloride Intake Chloride Output Chloride Absorption Chloride Retention of Period Age Weight per twenty—four per twenty—four per twenty—four per twenty—four hours Hours Hours Hours Total er Kg. Feces Urine Total Total Per Kg. % Total Per Kg. ,% Author Breast Milk Intgke Intake Days Mo. Kg. Gm. Gm. Gm. Gm. Gm. Gm Ga. Gm. Gm. Freund 1 4 4.5 4.94 0.170 0.034 0.07 0.126 0.1 3 0.163 0.030 96.1 .037 0.007 .21.8 2 6 3.0 2.95 0 238 0.081 0.07 0.156 0.1 3 0.231 0.078 97.0 .075 0.025 _25.4 Tobler L. I and 1 6 2.5 4.24 p.069 p.002 0.003 0.056 0.059 0.066 0.002 95.6 .010 0.000 14.4 I011 F. 1 2.5 4.28 0.254 0.059 0.01 0.1 2 0.16 0.241 0.056 94.2 0.088 0.021 ,34.8 4.37 0.255 0.0 6 0.02' 0.1 2 0.16 0.231 0.053 90. 0.089 0.020 ‘34.8 Lindberg G. 4.37 0.185 0.0 0.016 0.072 0.088 0.169 0.039 91.3 0.097 0.022 ‘52 4 4.37 0.195 0.044 0.036 0.086 0.122 0.159 0.036 81.7 0.073 -37 6 Blauberg 1 6 5 ——_- 0.203 ----- 0.014 0.013 0.027 0.189 ----- 93.1 0.176 _____ Michel C. and _-_ ——— 3 4.725 0.278 0.059 0.010 0.199 0.209 0.268 0.057 96.6 0.069 Ferret W. Klotz M. _—— _-— 2.5 3.42 0.224 0.066 0.073 0.056 0.130 0.151 0.044 67.2 0.094 Cows Milk Blauberg _—- -—— ——— —--- 0.173 ----- 0.071 0.239 0.310 0.102 ----- 58.9 If.138 ----- Tongle -__ _-_ 4. 1 0.890 0.197 0.029 0.791 0.81 .862 0.191 96. .0 1 3.7 5.21 0.696 P 129 0.025 0.610 0.633 .671 0.12 96.2 .0g1 * Calculated from the table giving the NaCl metabolism of healthy infants. (13 - h - eleven days and six months. The subject were fed alter- nately with breast milk and modified cows' milk for a period of six days - three of which were for adjustment to the diet and three eXperimental. Analysis were made of the chlorine content of food, feces and urine. Table II which gives a summary hf the studies reported in American literature reviews the results of Wang and Davis. Their data show that the chlorine intake of the babies on a diet of cows' milk exceeded their intake on breast milk, the average being 0.898 gram and 0.268 gram reapectively per twenty-four hours. The amount of Chlorine excreted in both feces and 'urine was greater when cows' milk was fed. The average excretion in the feces was 0.068 gram and 0.035 gram and in the urine 0.666 gram and 0.179 gram on the reSpective diets. The absorption of chlorides ran parallel to the urinary chloride. The average on the diet of cows milk was 92.4 'percent and on the breast milk $7.0 percent of the total intake. There was a great variation in the chloride retention of different subjects both in the percentage of the intake and in the amount Which was retained per kilogram of body ‘weight. There appeared to be no relation between the amount ingested and the amount retained. The average percentage retained on either kind of milk was approximately the same, being 20.6 percent for breast milk and 18.2 percent for TABLE II on er twenty—four hour e on per twenty—four Author e of Diet Period per twenty—four EGGS I18 er Hoobler (H) __________ 3 Wakeman, Shohl and Sato (9) 3 Bawman and - 6 - cows' milk. The authors found that as age increased, there was a decrease in the amount retained with cows milk, but that no such relation existed with the breast milk. No definite relation between age and percentage retention could be found. Shoh1,_Wakeman and Shorr (8) investigated the mineral metabolism of an eight months old male infant on a normal and on a high mineral diet. The study was divided into two periods. In the first period of four days the subject received a diet of skimmed cows' milk and cane sugar to which salts had been added to make the concentration of each mineral approximately double that of cows' milk. After a ten.day preliminary period, the same food, without the salt addition, was fed for five days. Excretion in the trrine and feces was determined and a balance of each of the ruinerals computed. The conclusions were that a baby on a skimmed milk (iiet, after salt constituents were doubled, retained salts in.approximately the same prOportion but in twice the amounts. However, the data submitted in Table II reveals ‘that the chlorine retained on the high mineral diet (0.80 gram) was approximately seven times greater than on the diet without: the added salts (0.11 gram). Shohl and Sato (9) studied the effect of acid and base on the metabolism of two seven months old infants for'two three day periods. The normal food consisted of I a... I)- .ICOEML'L .I. MIDAMA‘V In“; or...» TL... 1.. ' viva .. i - 7 - whole milk and water mixtures with added cane sugar, and was fed to each infant for one period. Then, without alteration of the diet, in one case 250 c.c. .1 N HCl were added,.and in the other, #73 0.0. .1 N NaHCO3. Food, urine, and feces were analyzed and the acid base value calculated. During the period of normal feeding the reactions of the two subjects were quite similar. On the kilogram basis, the infants eliminated 0.074 gram and 0.086 gram of chlorine respectively, while the absorption ranged from 0.088 to 0.098 gram for the two subjects. On an intake of approxi- mately 0.09 gram per kilogran the babies retained 0.015 and 0.013 gram. Though the intake during the acid administratbon was approximately double that of the normal and base feedings, a negative retention of -O.267 gran resulted. While a slight negative retention (-0.032 gram) occurred when sodium bicarbonate was fed the output and absorption consistently followed those of the normal period. In a series of metabolism studies Hoobler (h) fed both cows milk and breast milk to a nine months old infant. He found that the excretion of sodium chloride took place almbst entirely through the urine and that the absorption was 96 to 98 percent of the intake. Although the intake on cows' milk was greater than that on breast milk, the actual retention was greater on breast milk. The author, however, stated that the retention of sodium chloride was only suf- ficient to cause a balance to be maintained in body fluids under normal conditions. Sawyer and her coworkers (7) studied the mineral metabolism of two apparently normal male children aged five and eight years respectively. The investigation con- sisted of two three day periods during one of which the children received a normal and, on the other, a high fat diet. The mineral, calorie and nitr0gen intakes for the two diets were practically identical. The data, included in Table II, show that the intake per kiIOgram of body weight during the normal period was approximately 0.30 gram for both children. While the fecal chlorine varied the urinary chlorine was very similar, namely, 7.242 grams and 7.029 grams respectively. Both subjects absorbed practically 99 percent of the intake. ZDuring the period of normal feeding the two children showed negative retentions of -0.018 gram and -0.005 gram jper kilogram of body weight. On the high fat diet, however, one subject retained 0.016 gram per kilogram of body weight. This retention compares favorably with the results obtained by Shohl and Sato (9) and with some of the results found by Wang and Davis (12) On the infants fed breast milk. The only long time experiment found was that conducted by Clark (3) who studied the mineral metabolism of five men 111 the California state prison for a period of twenty-eight weeks. These men had no history of disease which might lead to abnormal metabolism. The experiment consisted of _ 9 - five periods in which the diet varied in the cereal, sugar, vegetable, and milk content. During the last period magnesium oxide and calcium lactate were given orally. The intake, output and retention were recorded in weekly periods both in grams per week and in c.c. of normal solution. Table III shows the average daily balances computed from these data which indicate that the amount of chlorine ingested per kilogram of body weight ranged from 0.062 to 0.094 gram for all of the subjects. The average daily chlorine retentions showed considerable variation for each period as well as in different periods for the same subject. When compared on the kilogran'basis, the variations in the retention for the different subjects correSponded to the differences in the intake. With some exceptions, the re- 'tentions for a given individual tended to increase as the diet was increased in its chlorine content. The data regarding chlorine metabolism obtained by ‘various scientists indicate that chlorine is excreted chiefly in the urine, that over 90 percent of it is absorbed, and.that, unless the amount ingested is too great, there is, 111 general, a higher retention per kilogram of body weight on a higher intake. The fact that on a similar intake infants and adults retained approximately the same amount of chlorine per kilogram of body weight may perhaps signify 'that the retention on this basis does not vary significantly 'with.age. However, a search of literature revealed no TABLE III S HYAHY OF CLARK'S CHLOHTHE EETABOIISH STUDIES ON HEN Chloride Intake Chloride Output Chloride Absorption Chloride Retention Subject Age Weight per twenty— per twenty—four per twenty—four per twenty-four four hours hours hours hours Total Per Feces Urine Total Total Per Percentage Total Per Percentage Kilogrgg. Kilogram of Intake ilogram of Intake 1.... gm Yrs Kg. Gms. Gms. Gus. Gms. Gms. Gms. Gms. Gus. Gms. J. w. 42 57.7 4.090 0.071 0.038 2.818 2.856 4.052 0.070 99.0 1.234 0.021 30.2 s. K. 22 62.3 5.400 0.087 0.146 4.302 4.448 5.254 0.084 97.2 0.952 0.015 17.6 L. M. 31 67.7 5.880 0.087 0.088 4.686 4.774 5.792 0.085 98.5 1.106 0.016 18.8 J. 8. 49 57.7 4.570 0.079 0.135 2.820 2.955 4.435 0.077 97.0 1.615 0.028 35.3 E. E. 36 80.9 5.510 0.068 0.065 5.430 5.495 5.445 0.067 98.8 0.065 0.008 1.2 \1 _ 10 .. - 11 _ record of long time studies of chlorine metabolism of normal preschool children. Sawyer and her coworkers (7) reported the only balance study made on apparently normal children. Unfortunately both of the subjects were in negative chlorine balance, and the results, therefore, cannot be compared directly with these reported in this study. Since almost all of the observations on the chlorine metabolism of infants were conducted for a short period of time' there is a need for further studies. Procedure The experiment reported here is a portion of a calorie, protein, and mineral metabolism study conducted on two preschool boys, D.A. and W.W. aged four years and :2 nine months and four years and seven months reapectively. Throughout the study the children lived under supervision in the Home Economics Building. According to physical and medical examinations made before this experiment, the children were in excellent physical condition. Table IV records the height and weight of the children and the percent variation of these from the Woodbury Height-Weight- Age Tables. TABLE IV THE HEIGHT AND WEIGHT 0F TEE CHILDREN AS COEPARED TO THE WOODBURY HEIGHT—WEIGHT—AGE TABLES (13). Weight He ght Subject Age bserved Normal fl bserved ormal % Var- ariation iation Ho. Lbs. Lbs. In. In. w.w. 55 41.75 36.47 .12.6 43.75 40.67 +7.0 D.A. 57 38.38 37.14 + 3.2 42.88 41.08 +4.2 According to age W. W. was 12.6 percent and D. A. 3.3 percent above the average weight. Their heights were 7.0 and 4.2 percent reSpectively above the average given for their age. Throughout the study the children lived under super- vision and were kept under a regular routine as to their meals, toilet,habits, sleep and outdoor exercise. The i I-m} :» 'L -13.. investigation extended from February 1 to March 22, 1932. After a six day preliminary period the study was divided into two parts - a thirty day period in which the diet furnished three grams of protein and a fifteen day period in which it furnished four grams of protein per kilogram of body weight. These were considered a medium and a high level of protein intake. This paper reports the chlorine balances during the last nine days of the medium protein level and the first twelve days of the high protein level. Tables V and VI give the diets for the two periods and indicate that the caloric content was approximately the same. Observation of these tables show that the milk, egg, butter, cod liver oil, vegetables and fruits included in both diets furnished an adequate supply of minerals and vitamins. All food which the children received was carefully weighed, and, in every case, all that was served was eaten. To insure an even sample the food was first mixed, then ground or pureed. Sufficient food for a three day period was prepared at one time and daily samples were weighed and stored in a refrigerator. The food was cooked and served in the same dish. A small amount of sodium chloride was added to the food for the first three day period on each diet and none thereafter. Four hundred grams of distilled water were used daily for drinking and for food preparation. Duplicate samples of food, equivalent to the amount D. A. received, were saved for analysis. These were dried, sH mHepos -unuw 11:» us: nu. H lwmw Hao uw>Ha so 3...... .5-.. M86 9H. H We Ianm them Imam {in m .2... drum 1... mHml 2. b magnum or act: .400. Ho. II: o H oo omsHeHWI s.OH. 0Ho.q1:mMeo.m m m.o sH mosppwm m.os” :00. o 0H0. mm M40 om» mossmmHnm4 smchshpm M o..m .m o. m o. m.o ow museum emeHsHpm _ o . o. m.o N muonneo pesHmem . filo.o 00 H H om weepssos emehsnpm m.:mn m N emo.o me.Q NW l smv. mo opepom emxbmm . . m Ho.o «Ho.o .4m H seHmam saxoooep it: o.mm mmwuo lemma sWH H mN 33m page 39: . . m .. . o. H.0H w. meom tam ._ mam: . So. m mLLm I} m. m.mmH w .0 new. ofi m: m.mm owe MHHa mHoe: . onz oo 0H\z oo mmwm oHo< .maw .mew .msm .mem 0: Heme maHMOHao esflsom mmHHOHao qusosm seaHme swim HH 0H m m emHo szeomm mpHmms we we onsHmomsoo > mqm¢a :l’lfl'in F1|§3vll an. . 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Daily collection of feces were also made using charcoal and carmine alternately as markers. These were dried and stored in the same manner as the food. In the eXperiment the analysis were made by the Peters and.VanSlykes (5) modification of the Volkard-Harvey titration. The ferric alum,silver nitrate and nitric acid were combined into one solution. This combination made one less reagent solution and the nitric acid stabilized the Agn03. To insure a more permanent and point, the silver chloride was filtered off before titrating with ammonium sulfo cyanate. The accuracy of the method was tested by recovering chlorine from known solutions. These data, as shown in Table VII, indicated that the recovery was within 2 percent of the theoretical value. The food and feces were ashed in an alkaline medium (1). From 3-5 cc of 5 percent NAQCO} were added to the weighed samples of food or feces and evaporated to dryness on a hot plate. The samples were then ashed in an electric furnace at a full red heat. The ashed samples were taken up in hot water. The filter paper and residue were then ashed, and taken up again in lzh ENC}. The entire ashed sample was used for a single analysis. Determination of urinary chlorine was made on urine samples previously preserved with toluene. Either duplicate or triplicate. determinations were made in all case s. -17.. TABLE v11 RESULTS or TESTS TO SHOW THE ACCURACY OF THE CHLOEINEiHETHQD Urine K01 Trial __ Chlorine Present Calculated Analyzed"I Percentage Recovered cc cc gm. gm. 2 1. 0.075 0.0744 99.1 2 0.0740 98.6 3 0.0743 99.1 0.0742 98.8 5 0.0745 99.3 10 1 0.017 2 0.017 a 0.0176 0.0174 5 0.0177 5 2 1 0.0925 0.0910 98.2 2 0.0911 98.3 a 0.0915 98.8 0.091; 98.6 5 0.09 99. A: :AII— 1 ‘ Average of three trials. Discussion 2: Results Table VIII records the results of the daily cHorine metabolism for the last nine days on the medium protein diet. Since the food was weighed in three day periods these data have also been calculated on this basis and are given in Table IX. These two tables will be discussed at the same time. The daily chlorine intake during the first 3 day period was 2.353 grams for D. A. and 2.588 grams for W. W. These values were practically 0.40 gram higher than those of the other two periods because a small amount of sodium chloride was added to the food. The average intake was 2.120 grams for D. A. and 2;313 grams for W. W. The total percentage of the chlorine excreted ranged from 83 to 86 percent of the total intake. In both sub- jects practically all of the chlorine (80 and 83 percent) was eliminated by the way of the kidney. The urinary output of D. A. followed closely the variations in the intake. 0n the first day the excretion was low. During the next three days the highest elimination of chlorine occurred and corresponded, with a one day lag, to the period when the chlorine was highest in the diet. These values were comparatively constant during the remainder of the medium protein diet. W. W. showed a greater daily fluctuation in the urinary chloride than did D. A. Nevertheless, with the -18.. .ImH' .EM 335 .30 poo 33 HHHb Hamds NH HA9; -21.. exception of the third day, the results showed the same tendency, to decrease on the day following a decrease in the chlorine content of the diet. The average urinary excretion was 1.691 grams for D.A. and 1.916 grams for W.W. When these data were calculated on the three day basis (Table IX) the output was quite constant regardless of the intake. The average fecal chloride was practically identical for the two children, namely 0.060 gram for D.A. and 0.059 gram for W.W. The amount of chlorine minus that which appeared in the feces was considered absorbed. Because of the high solubility of chlorine each child absorbed an average of 97 percent of his intake, and none of the values fluctuated far from this average. The grams of chlorine absaflrfl.were directly preportional to the intake in the case of each child. These results agree with those of Hoobler (4), who, in his study on infants, stated that the chlorine absorption was usually over 90 percent of the intake. The retentions for both subjects varied greatly. The values for D.A. were greatest during the first three days which corresponded to the period of the highest chlorine intake. There was a lowered chlorine retention on the fourth day, but the remaining five days showed a tendency to become constant. This tendency is also shown when these data are averaged in three day periods. The two highest balances for W.W. occurred on the first and third days when -22- the chlorine content of the diet was highest. 0n the fourth day of this eXperiment W.W. showed a negative re- tention which was preceded by an unusually high retention. This negative balance may have been due to the elimination of chlorine held over from the high retention of the day before. The average retentions as calculated by three day periods also Show that the most chlorine was retained during the period of highest intake. The average retentions were 0.363 gram for D.A. and 0.337 gram for W.W. The percentage of the intake retained for both children varied from 3 to 39 percent. The data show definitely that there is a greater percentage as well as a greater number of grams of the chlorine retained with an increased intake. The average percentage of the intake retained by D.A. and W.W. was 17.5 and lfl.6 percent reapectively. Table X gives the chlorine metabolism of the two children on a medium protein diet expressed in grams per kilogram of body weight. The chlorine intake during the first three day period was 0.135 gram. The intake for the next six days was practically constant furnishing approxi- mately 0.11 gram per kilogram. The respective averages for the total output, 0.100 and 0.104 gram were practically identical for the two children. These data show the same tendencies as those discussed under the total chlorine, namely: excretion of -23.. TABLE X THE DAILY CHLORIFE HETABCLISH ON A MEDIUM PROTEIN DIET EX— tention gms. gms. - 2h - practically all of the chlorine through the kidney, an increased excretion of this element in the urine on the day following an increase in the diet, a fairly constant fecal output and a variation in the total elimination from day to day. The daily absorption on this basis quite closely followed the variations in the intake. Based on average results D.A. absorbed 0.118 gram and T.W. 0.119 gram per k110gram of body weight. The greater retentions (with the exception of the second day for W.W.) occurred when the chlorine intake was the highest. It is interesting to note that the one negative retention for W.W. and the lowest retention for D.A. were an the first day following a reduction of chlorine in the diet. The average retentions per kilogram of body weight were approximately the same being .021 gram for D.A. and .018 gram for W.W. The data per kilogram of body weight calculated as average figures per three day periods is shown in Table XI. This table reveals clearly the similarity between the reaction of the two children. The average amount of the intake absorbed was virtually the same for both subjects. The average urinary elimination of D.A., being approximately 3 percent less than that of W.W., caused his average retention to exceed W.W.'s by that amount. Table XII records the data for the daily chlorine -25- TABLE XI THREE DAY AVERAGE OF THE DAILY CHLORINE METABOLISM ON A MEDIUM PROTEIN DIET EXPRESSED IN GRAMS PER KILOGRAM OF BODY e 0d on .E& waquH sen sooHnsm HHN Hamda . . o . . . u a I u o s o D 0 o c 0 u p o n V . . o o t o a o v 0 I c 0 o a 9 O 0 o c I 0 - I C v a 0 V c I . O u o n c D O a v I c . v . o v a v . . . a a . v v . . . . . . . . . . - o c o 0 o . I . . . . . . o c . . v . o - u u . o q u c I P h o l . . . . . . . o . A o n _ o v n I - a — o u a o 1 1 . y . q s v f c t O 0 o . n I a g o n 0 O o b v r r . p u n v r v d o o O o . o a I . . 0 . . o r 5 n c . s p u u p . n g . . u a i u o a u a u 0 u :1 lulu? ’Ibilhlli .l. w5.\.l.l ll. .. h v -27.. Inetabolism during the first twelve days of the high protein diet. Table XIII which gives these values averaged by three day periods will also be discussed at this time. The diet during the first three day period, the chlorine content of which was increased approximately one gram over the last three day period of the medium.protein diet, contained a lrigher amount of chlorine than any other period. During the next two periods of six days the chlorine intake was con- stant and was practically 0.5 gram lower than during the first three days. On the last three days the chlorine content of the diet was lowered 0.2 gram below that of the 'previous period. The average chlorine intake for D.A. was 2.499 grams and for W.W., 2.749 grams. The daily urinary chloride varied from 2.358 grams to 1.730 grams for D.A., and from 2.626 grams to 1.986 grams for W.W. The highest urinary output for D.A. occurred on the same days that the chlorine intake was the highest. With two exceptions the output followed the fluctuations in the intake and there was no evidence of a one day lag as was revealed on the medium protein diet. The data for W.W. do not show such a clear cut tendency to follow the intake when the daily values are considered, but if the values are averaged into three day periods (Table XIII) there is again a relationship between intake and output. The daily fecal output of chlorine ranged from 0.025 to 0.086 gram, the average being 0.046 gram for D.A. and 0.055 gram for W.W. These average values are almost 00H sous HHHN Ham»; -29.. identical with those on the medium protein diet. The total percentage of the chlorine excreted ranged from 76 to 92 percent of the intake - 80 to 82 percent in the urine and 2 percent in the feces. It is interesting to note (Table XIII) that the average percentage of the chlorine excreted in the urine was practically the same as that on the medium protein diet, namely, 80 percent for D.A. and 82 percent for W.W. Although the urinary values are less consistent on the high protein than on the medium protein diet, the daily chlorine absorption was more constant, and in each case it followed the chlorine intake closely. Therefore, more grams of chlorine were actually absorbed by the kidney. The absorption ranged from 97.1 to 98.7 percent for D.A. and from 96.8 to 99.1 percent for W.W. The average daily absorption for the two children however, was the same and it was less than 1 percent higher than the average absorption on the medium protein diet. Again W.W. showed a greater variation than D.A. in the amount of chlorine retained, though both children again showed a tendency to store more chlorine on the days when the intake of this element was the greatest. This fact (Table XIII) is eSpecially evident when the data are averaged in three day periods. The average retention for D.A. was 0.447 gram and for 8.8. 0.438 gram. Each child retained almost the same percentage of his chlorine intake whether he had a high or a medium protein - 3o - diet. The average values were 17.5 and 17.9 percent for D.A., and l#.6 and 15.9 percent for W.W. These data also show that there was only a 2 to 3 percent difference in the amounts retained by the two children. Table XIV records the daily chlorine metabolism on a high protein diet eXpressed in grams per kilogram of body weight. Table XV gives these data averaged in three day periods. The chlorine intake varied from 0.13 gram to 0.17 gram for either child. As previously discussed the chloride excreted in the urine generally followed the changes in the intake. The average values excreted in grams per kilogram of body weight were 0.115 gram for D.A. and 0.119 gram for W.W. These amounts were slightly higher than those on the medium protein diet, but increased in proportion to the increase in diet. The amount of fecal chlorine on this basis was the same for the two children (0.003 gram) and was fairly constant day by day. The average fecal chlorine per kilogram was the same on the high as on the medium protein diet for both subjects. The daily absorption followed the intake very closely, the average value being practically identical for the two children (0.1Ml and 0.142 gram). The body actually absorbed more grams as well as a greater percentage of the chlorine intake on the high protein than on the medium protein diet. Approximately 98 percent of the intake was absorbed in both cases which is a value higher than those obtained by the majority of early investigators. Some -31.. TABLE XIV DAILY CHLORINE METABOLISM ON A HIGH PROTEIN DIET EXPRESSED W.W. 18.98 IF.I|'II vb! t! ‘I. L. -32.. TABLE XV THREE DAY AVERAGE OF THE DAILY CHLORINE METABOLISM ON A .3IGH PROTEIN DIET EXPRESSED IN GRAHS PER KILOGRAM OF BODY. - 33 - American workers (Hoobler (U), Shohl and his coworkers (8), and Sawyer, Bauman and Stevens (7)), however, ob— tained absorption figures which were correSpondingly high. The average retentions per kilogram of body weight were approximately the same for both children. 0n the high protein diet, which furnished 0.1h-gram of chlorine per kilogram of body weight, D.A. and W.W. retained 0.026 and 0.023 gram reSpectively. These values were very similar to the 0.021 and 0.018 gram retained on the medium protein diet which furnished 0.12 gram of chlorine. The results of foreign investigators, as shown in Tabb I, indicate that on the kilogram basis the chlorine intakes were lower and usually resulted in lower retentions than those in the experiment reported here, although the percentages of the intakes retained were similar. In the cases where the intakes were higher, the retentions were usually proportionally higher which indicates, as does this experiment, that there is some relationship between intake and retention. 0n the other hand, on a greater chlorine intake (0.30 gram) Sawyer and her coworkers found that their subjects were in slight negative balances (—0.018 and -0.005 gram per kilogram of body weight). There is a possibility, however, that these subjects may have been accustomed to a higher chlorine intake previous to the experiment. The adult subjects studied by Clark (3) received a slightly lower intake per kilogram than did the children in -324- this study, but they retained approximately the same mount of chlorine. This substantiates the opinion of Wang and Davis (12) that possibly there is no relation between age and retention. Definite conclusion regarding the amount of chlorine needed by pre-school children can be drawn neither from the literature cited nor from this study. The result of this investigation reveal, however, that with the exception of one day, the children were able to remain in a positive chlorine balance over a period of twenty-one days on an intake as low as 0.12 to 0.14 gram per kilogram. This indicates that probably they were receiving a sufficient amount of this element in the diet. Since very little sodium chloride was added to the diet, these results appear to confirm the theory that very little, if any, tabhe salt need be added to the food of preschool children. SUB-{MARY *- A study of the chlorine metabolism of two normal preschool boys fed either a medium or a high protein diet was conducted for a period of twenty-one days. 1. The average intakes on the medium diet were 2.120 gramsfor D.A. and 2.313 grams for W.W. while those on the high protein diet were 2.499 grams and 2.7u9 grams respectively. With the exception of one day for one subject the children were in a positive balance throughout the entire study. From 80 to 83 percent of the chlorine intake was ex- creted in the urine. The fecal chloride was identical for both subjects on both diets averaging from 2 to 3 percent of the intake. 0n the kilogram basis the total amount of chlorine ex- creted by the two children was practically the same. The daily absorption followed the intake closely, average values ranging from 97 to 98 percent of the ingested chlorine. The average amounts of chlorine retained per kilOgram of body weight on both diets were similar for the two subjects, namely 0.021 and 0.026 gram for D.A. and 0.018 and 0.023 gram for W.W. -25... 8. 9. -36- The fact that the high protein diet furnished slightly more chlorine than the medium protein diet did not appear to materially affect the percentage of the intake absorbed or retained, but did cause an actual increase in the number of grams retained. No definite conclusions can be drawn regarding the amount of chlorine needed by preschool children. APPENDIX -35.. TABLE XVI DAILY CHLORINE METABOLISM ON A MEDIUM PROTEIN DIET EXPRESS‘ gm. @1- Sm- gm. gm. -33- TABLE vaI 1 THE DAILY CHLORINE METABOLISM on A HIGH PROTEIN DIET rx— 2. 30 10. 11. 12. 13. 1 ,_ BIBLIOGRAPHY Association of Official Agricultural Chemistry, 3rd Ed., Chap. 12, Page 111 - 1930. Blatherwick, N.R.,: Studies of Urinary Acidity II The Increased Acidity Produced by Eatin Prunes and Cranberries. J.B.C. 57, pp. 815-81 , 1923. Clark, G.W.,: Studies in the Mineral Metabolism of § Adult Man. University of California Publica- ; tions in Physiology 5, #17, pp. 2M7-5l, 1926. Hoobler, R.B.,: The Role of Mineral Salts in the r Metabolism of Infants. Amer.Jour.Dis.Child. ’ 2, 106-lu0, 1911. , Peters and VanSkae,: Quantitative Clinical Chemistry Methods. p. 833-835, 1932 (Williams and Wilkins). Rose, Mary S.,: Handbook for Dietetics, 3rd Edition, 1929 (Macmillan). Sawyer, Margaret, Bauman, L., and Stevens, F.,: Studies .1 of Acid Production. II The Mineral Loss During r Acidosis. J.B.C. 33, p. 103-109, 1918. ' Shohl, A.T., Wakeman, A.M., and Shorr, E.G.,: Mineral Metabolism on a High Mineral Diet. Amer.Jour. . Dis.Child. 35, p. 576-579, 1928. ' a Shohl, A.T., and Sato,: .Acid Base Metabolism I. Determination of Base Balance. J.B.C. 58 p. 235-253, 1923-2u. ‘ , Storms, Lillian B.,: Analytical Data Included in Pamphlet I'Babys' Vegetables - Gerber Products Co., Fremont, Michigan. Waller, Dorothy S.,: Nutritive Value of Foods. George Wahr, Ann Arbor, Michigan, 1932. Wang, Chi Che, and Davis, L.H.,: A Comparison of the Metabolism of Some Mineral Constituents of Cow's Milk and of Breast Milk in the Same Infant. Amer.Jour.Dis.Child. 27, p. 569-77, 192k. Woodbury, R.M.,: Heights and Weights of Children Under Six Years of Age. U.S.Department of Labor, ChildrenS' Bureau #87 Table xxx1,p. 76. "154' - "IAlllitill‘i‘ll'lilfllfl